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Influence of heat-labile serum components in the presence of OmpA on the outer membrane of Salmonella gallinarum

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Abstract

Salmonella gallinarum is the causative agent of fowl typhoid. Being a Gram-negative bacteria, its outer membrane proteins (OMP) can be regulated by different microenvironments. S. gallinarum was cultured under the following conditions: nutrient broth (NB), NB supplemented with serum from specific pathogen-free birds (NBS) and NB with serum incubated at 56 °C prior to incubation with the bacteria (NBSD); OMP were subsequently extracted. Several changes were observed in the apparent expression of OMP, mainly a decrease in an OMP with a size of 30 kDa, approximately, under the NBS condition. In contrast, the same event was not observed in NB and NBSD when using one- and two-dimensional polyacrylamide gels (SDS-PAGE). Using the OMP with a size of 30 kDa, approximately, as antigen in indirect ELISA, we were able to differentiate serum from healthy and vaccinated birds, as well as birds infected with S. gallinarum and S. enteritidis. The amino-terminal of this protein was sequenced, showing 100 % identity with OmpA of S. typhimurium. Subsequently, we designed primers to amplify the gene by PCR. The partial sequence of the amplified gene showed 100 % identity with OmpA of S. gallinarum. (1) Heat-labile serum components influence the presence of OmpA in the OM of S. gallinarum; (2) by the way of ELISA, OmpA allows to specifically differentiate healthy from diseased birds.

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Acknowledgments

In memoriam of Dr. Guilllermo Mendoza Hernández for the sequencing of the protein amino-terminal ending. Facultad de Medicina, UNAM. We thank CONACyT and DGAPA (Universidad Nacional Autónoma de México) for their financial support by projects CONACyT 27669-B and PAPIIT IN223398, respectively.

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Authors and Affiliations

  1. Facultad de Agronomía y Veterinaria, Universidad Autónoma de San Luis Potosí, San Luis Potosí, SLP, Mexico

    X. Vega-Manriquez

  2. Laboratorio de Microbiología Molecular, Departamento de Microbiología e Inmunología, Facultad de Medicina Veterinaria y Zootencnia, Universidad Nacional Autónoma de México, Circuito exterior S/N, Ciudad Universitaria, UNAM CU, Coyoacan México, 04510, México City, Mexico

    L. Huerta-Ascencio & A. Verdugo-Rodríguez

  3. Departamento de Producción Agrícola, Universidad Autónoma Metropolitana-Unidad Xochimilco, México City, Mexico

    D. Martínez-Gómez

  4. Programa de Inmunología Molecular Microbiana, Facultad de Medicina, Universidad Nacional Autónoma de México, México City, Mexico

    Y. López-Vidal

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  1. X. Vega-Manriquez
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  2. L. Huerta-Ascencio
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  3. D. Martínez-Gómez
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  4. Y. López-Vidal
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  5. A. Verdugo-Rodríguez
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Correspondence to A. Verdugo-Rodríguez.

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Communicated by Jorge Membrillo-Hernández.

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Vega-Manriquez, X., Huerta-Ascencio, L., Martínez-Gómez, D. et al. Influence of heat-labile serum components in the presence of OmpA on the outer membrane of Salmonella gallinarum. Arch Microbiol 198, 161–169 (2016). https://doi.org/10.1007/s00203-015-1174-4

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  • DOI: https://doi.org/10.1007/s00203-015-1174-4

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